Choriogenin mRNA induction in male medaka, Oryzias latipes as a biomarker of endocrine disruption

Abstract

The fish egg envelope has been known to consist of two distinct layers. The outer layer is thin and is formed around the oocytes during the later stage of previtellogenic development. The inner layer, called zona radiata, is thicker than outer layer and occupies most of the egg envelope. Zona radiata proteins of medaka (Oryzias latipes) consist of two major subunit groups, ZI-1,2 and ZI-3. The precursors of ZI-1,2 and ZI-3 have been named choriogenin H and choriogenin L, respectively. They are synthesized in the liver in response to estrogen, and then released into the blood stream and incorporated into the zona radiata in sexually matured female medaka. However, choriogenin is also induced in male medaka when the fish are exposed to estrogenic chemicals. Choriogenin can be suggested as a possible biomarker of endocrine disruption in fish, however, relatively little has been known about chorogenin gene expression by estrogenic chemicals. In this study, full sequence of choriogenin L DNA was identified, and measurement of choriogenin mRNA induction was established in medaka by use of RT-PCR technique. Also estrogenic effects of industrial chemicals were evaluated by this tool. When mature male medaka was treated with bisphenol A, nonylphenol, and 17alpha-ethinylestradiol for 6 days, respectively, the expression levels of choriogenin L and H mRNA were elevated in treated groups. The induction of choriogenin subunits expression by these chemicals showed a dose-dependent pattern and choriogenin L was found to be more sensitive than choriogenin H.